AU6232080A - Cyclohexane carboxylic acid derivatives - Google Patents

Description

COIMONWEALTH OF AUSTRALIA

Patents Act 1952-1979 ~LLA i'iICATION FOR A STANDARD P

62320 2-chome, NIPPOCEEMIPHAR CO., LTD. of No. 2-3, Iwamoto-cho, nuiyoda-ku, Tokyo, Japan and' TEIKOKU CHEMICAL INDUSTRY CO., LTD. Of No. 1-18, Kita-Hori--oef r"IlPishi-ku, Osaka-shi, Osaka-ft, Japan two companies organized and existing under the laws of Japan hereby apply for the grant of a Standard Patent for an invention entitled

"1CYCLOHEXAWE CARBOXYLIC ACID DERIVATIVES"

h~isdescribed in the accompanying complete specification.

SSEP 198o DETAILS OP BASIC APPLICATION(S)::- Nt""b0 f bas~ic application:- 54-120142 54-168271 54-168272 Name of Convention country in which basic application was filed:- JAPAN JAPAN JAPAN Date of basic application:- 20th September, 1979 26th December, 1979 26th December, 1979 00Our address for service is: F.B. RICE CO., 101 Mort St, Balmain 2041

Dated this 10th day of September 1980.

INUSR Co

TEIOKUCHEMIC D

By:

TO: The Commissioner of Patents F.B. RICE CO., CObBIONEALTH OF AUSTRALIA Patent Attorneys, Sydney.

Farms 7/8 COMMONWEALTH OF AUSTRALIA Patents Act 1952

DE CLAIRATION IN SUPPORT OF AN~ APPLICATION-QR-- A CONVENTION APPLICATION FOR A PATENT OR P AT T OFlqrADDT- T 62320/80 In support of the Appli(;atiQn made by Convention Application NIPPON CHEMIPHAR CO., LTD. and TEIKOKU CHEMICAL INDUSTRY CO., LTD. for a paet -for an invention entitled: patent of addition CYCLOHEXANE CARBOXYLIC ACID DERIVATIVES IAlIRA YAMAGUCH1I President, and ETNOSUKE NAGASE President of 000S and care of the applicant companies Ce: of and care of the applicant company do soleminly and sincerely declare as follows:

*e forth~ patent *patent ot additi-on

1 am authorised by the applicant for thte to make this declaration C* on its behalf.th.

2 The basic application(s)l as defined byv section L_ of the Act made 142 were iJapan on20th Septer~oer, 1979 in pa 26hDeebr,17 Japan on 26th Decemnber, 1979) eecin on by NIPPON CHENIPFAR CO., LTD and TEIKOKU CHEMICAL INDUSTRY C e s* LTD.

(see back hereof)

is/are the actual inventor(s) of the invention and the facts upon which

U the applicant companies is/are entitled to make the application ar6 a flos. 0 The applicants are the assignees of the invention fromt the said actual inventors.

-b( 4 The basic application(.) referred to in paragraph 2 of this Declaration is/are the first application(s) made in a Convention country in respect of the invention the subject of the application.

Derlared at Tokyo this 29th day of August 19 Ro.

To: The Commissioner of Patents, 1. Commonwealth of Australia. Akrcata Cheyd phar Nippon i sdn P o ,Ltd.

This Fornm issuitable for any type of Patent Application. E.foueNgsPeie~ Status Teikoru Chemical Industjy Co-,.L~t NP legalisatFon required. d- F.B. RICE CO., -a Delete whichever is in-applicable. Patent Attorneys, nlet if not C(on,ention application, c-rr ly X~U I

(12) PATENT ABSTRACT (11) AU-A1-62 320/80 f(19) 0 TII'ATLI Ll (1 AU I_ (54) GUANIDINE-CYCLOHEXANE COMPOUNDS (71) NIIPPON CHEMIPHAR CO. LTD. (21) 62,320/80 (22) 11.9.80 (24) 20.9.79 (31) 54-120142 (32) 20.9.79 (33) JP 54-168271 26.12.79 JP 54-168272 26.12.79 JP (43) 26.3.81 (51)3 CO7C i29/12 A61K 31/22 A61K 31/44 A61K 31/355 C07D 213/63 CO7D 213/65 (72) Muramatsu, M. ET AL (74) RI 57) One object of the present invention is to provide novel

cyclohexane carboxylic acid derivatives which exhibit strong inhibitory effects on protease, anti-ulcer effects, anti-histamine effects, anti- inflammatory effects and anti-allergic effects. Claim

1. A compound of the formula:

-NHCH.2 a COOR 1 H2N

wherein R1 represenlts wherein Ri represents aa vanilyl, R2 pyridyl vanilyl, napthyl, napthyl, pyridyl or or ct-tocopheryl a-tocpheryl.

group, or a group of the formula, wherein R2 represents

a hydrogen atom, a lower alkoxy, formyl, lower alkanoyl or phenyl

group, or a q-Qup of the formula, -(CH 2)niCXOR, wherein R3 represents a hydrogen atom, a lower alkyl,,phenyl, benzyl, anisyl

120 ND 10.6

(21.1) AU-A-62,320/80 2 or 1owyr alkoxycarbcnylnwthyl group, and n represents an integer

of 0 to 2, or a pharmaceutically acceptable salt thereof. An anti-ulcer agent w.hich comprises a compound of the formula,

-NC2O O, C COOR3

wherein R3 represents a hydrogen atomor a. lowerx alkyl, phenyl, benzyl, anisyl or lower alkoxycarbonylmethyl group, and n represents an integer of 0 to 2,or a pharmaceutically acceptable salt thereof.

COMMONWEALTH OF AUSTRALIA

Patent Act 1952

COMPLETE SPEC I F I CATION (ORIGINAL)

Class Tht-. Class Application Number Lodged

Complete Specification Lodged 21.NOV1980 Accepted Published a4afS n49,

Priorities: 20th September, 1979 ees 26th December, 1979 d 3 a ft

26th December, 1979

Related Art .s *s i e 60 Name of Applicant NIPPON CHEMIPHAR CO., LTD. and TEIKOKU CHEMICAL INDUSTRY CO., LTD.

Address of Applicant No. 2-3, Iwamoto-cho, 2-chome, Chiyoda-ku, Tokyo, JAPAN

No. 1-18, Kita-Horie 1-chom', Nishi-ku, Osaka-shi, Osaka-fu, JAPAN

Actual Inventor MUTSUMI MURAMATSU TOSHIO SATOH YUKIO YANAGIMOTO ITADAMI SHINUCHI TOSHIO NAKAJIMA SISAO NAKAJIMA

Address for Service F.B. RICE CO., AT SUB-OFFJCE Patent Attorneys, ODGE The Forth and Clyde, SEP 1980 101 Mort Street, BALMAIN. 2041. Sydney Complete Specification for the invention entitled: "CYCLOHEXANE CARBOXYLIC ACID DERIVATIVES"

The following ,statement is a full description of this invention including the best method'of performing it known to us:-

BACKGROUND OF THE INVENTION

Field of the Invention:

This invention relates to novel cyclohexane carboxylic acid

derivatives, a process for producing the same and a pharmaceutical

Scomposition containing the same.

Description of the Prior Art:

Tranexamic acid (trans-4-aminomethylcyclohexane carboxylic acid)

is a cyclohexane carboxylic acid derivative, and is known to possess

excellent anti-plasmin effects. The esters of tranexamic acid are

also known to posss excellent anti-plasmin effects Okano et al,

J. Med. Chem., Vol. 15, No. 3, 247 (1972)). However, it was reported

that 4-guanidinomethylcyclohexanecarboxylic acid exhibited little e

anti-plasmin effects (ibid.).

A variety of cylcohexane derivatives has now been studied,

resulting in the present discovery.

SUMMARY OF THE INVENTION

One object of the present invention is to provide novel

cyclohexane carboxylic acid derivatives which exhibit strong inhibitory

effects on protease, anti-ulcer effects, anti-histamine effects, anti-

0 inflammatory effects and anti-allergic effects.

It is another object of the present invention to provide a

process for producing these novel cyclnhexane carboxylic acid

derivatives.

Further, it is another object of the present invention to

provide a cyclohexane carboxylic acid derivative useful as an anti-

ulcer agent.

These and other objects of the invention as hereinafter will

become more readily apparent can/be attained by the discovery of

compounds of the formula

HN

C-NHCH 2 COO (i) J

H2 N wherein R 1 represents vanilyl, naphthyl, pyridyl, a-tocopheryl R2 a group of the formula, wherein R 2 represents hydrogen,

•00 lower akoxy, formyl, lower alkanoyl or phenyl or a group of the

formula -(CH12 )nCOOR 3 wherein R3 represents hydrogen, lower alkyl,

phenyl, benzyl, anisyl or lower alkoxycarbonylmethyl, and n represents

an integer of 0 to 2. These derivatives and pharmaceutically

Sacceptable salts thereof have been found to possess excellent

inhibitory effects on protease, anti-ulcer effects and anti-

histamine effects, anti-inflammatory effects and anti-allergic

effects. Particularly, the compounds of the formula (II):

HN (CH2 )nCOOR 3

C-NHCH 2 (-C00 CII)

H2N

wherein R3 and n are the same as defined above, have been found to

S. possess excellent inhibitory effects on. gastric secretion, preventive and healing effects on various gastric and duodenal ulcers.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The ester residues R1 of the present compounds of the formula may be vaniyl, naphthyl, pyridyl, ct-tocopheryl or a group of R2 the formula /\Suitable naphthyl groups include ct-naphthyl

o *or f-aphthyl groups. Suitable pyridyl groups include 2-pyridyl, R

3-pyridJ]. or 4-pyridyl groups. Suitable groups of the formula

iniclude 6henyl, methoxyphenyl, ethoxyphenyl, propoxyphenyl,

fomnfyphenyl, acetylphenyl, propanoylphenyl, Dutyrylphenyl, biphenlyl or a group of the formula (H)CO 3 uiable groups

(CH cOOR .of the formula 2 n 3 include hydroxycarbonyiphenyl,

i,thoxycarbonylphenyl, ethoxycarbonyiphenyl, t-but.oxycarbonylphenyi, phienoxycarbonylphenyl, benzyioxycarbonylphenyl anisyloxycarbonyiphenyl, (e-thoxycarbonyl )methoxycarbonylphenyl. hydroxycarbonylmethylpheny3, methoxycarbonylmethyl pheny, ethoxycarboriylmethyl phenyl, t-butoxy- carbonylmethylphenyl, phenoxycarbonylmetiiylphenyl, benzyloxycarbonyl- methylphenyl, hydroxycarbonylethylphenyl, ethoxycarbonylethylphenyl, (ethioxycarbonyl )ethoxycarbonylethylphenyi, phenoxycarbonylethylphenyl,

benzyloxycarbonylethylphenyl, anisyloxycarbonylethylphenyl or the 0i

I

Compounds of the 'ormula may be either the cis~- or trans- isomer. Particularly preferable is the trans isomer. Thle pharmaceutically acceptable salts of the present compounds are the acid addition salts formed froim hydrochloric acid, sulfuric aci-d, phiosphcric acid, hydrobromic acid, acetic acid, lactic acid, maleic acid, fismaric acid, tIartaric acid, citric acid, methanesulfontc 4' 6

acid, p-Loluenesulfonic acid or the 1lke.

to£ According to the present invention, the compounds of the

*4 formula are produced by reacting 4-guanidinomethyl cyclohexane carboxylic acid or, a reactive derivative theareof with a compound of the formula (III): 1

R'I- OH (I

wherein RI I represents vanilyl, naphthyl, pyridyl, a-tocopheryl

or a group of the formula wherein R2' represents hydrogen,

lower alkoxy, formyl, lower alka-noyl, phenyl or a group of the

formula (cH2)nCOOR3', wherein F.3 represents lower alkyl, phenLyl, or loQJ6r o~iy~aymby ,benzyl,e-r anisy~, and n represents an integer of 0 to 2; and

when desired, removing the benzyl, anisyl or lower alkOxycarbonyl-

methyl group from the product.

Suitable reactive derivatives of 4-guanidinomethylcyclohexane-

carboxylic acid include the acid halides, for example, acid

chloride, acid bromide or the like, and mixed anhydrides using

ethyl chlorformate, butyl chlorformate or the like. Acid

halides are produced by reacting 4-guanidinomethyl-

cyclohexanecarboxylic acid with halogenation reagents such as thionyl chloride and thionyl bromide at a temperature of from room temperature to the boiling point of the halogenation reagent. The thus obtained acid halides are reacted with the compounds of the formula (III) to give the present compounds. This reaction is carried out by stirring at a temperature of from room temperature

'to the boiling point of the solvent for 1 to 40 hours. Suitable

which may be used include dimethylformamide, dimethylacetamide, aolvents

pyridine, dichloromethane, dichloroethane, chloroform, acetonitrile

or the like. Use of an acid-binding agent, triethylamine,e-e r pyrie dimethylaniline, is sometimes recommendable.

When 4-guanidinomethylcyclohexanecarboxylic acid is reacted

directly without conversion to reactive intermediate thereof, the

reaction is preferably carried out in the presence of a condensing

agent, for example, carbodiimide such as dicyclohexylcarbodiimide or

l-ethyl-3-(3' -dimethylaminopropyl )carbodiimide hydrochloride, sulfuric

acid-boric acid, carbodiimidazole, sufodiimidazole, or Lewis acid such

as phosphonousoxychloride or boron.trifluoride. The reaction is

carried out with a solvent, for example, toluene, xylene or dimethyl-

formamide, a solvent as mentioned above, or mixtures thereof at a

Lo temperature of from room temperature to the boiling point of the *aet o xmlcabdiiesc sdcylhxlabdiieo solvent.

The compounds of the formula wherein R3 represents a

hydrogen atom are produced by hydrogenating the compounds of the

formula wherein R3 represents a benzyll anisyl eeed-a1key-

-eatbo hey4mehy4 group in the presence of a catalyst, for example,

so*/1

-7-7

palladium. These compounds may also be produced by hydrolyzing the or loer JkoHxycarbonybwtyL compounds of the formula wherein R3 represents a 'butyllgroup

in the presence of a catalyst, for example, trifluoroacetic acid

or hydrochloric-acetic acid.

Compounds of the formula (IV)

HN S NHCH COO (IV) *C C

wherein R3 and n are the same as defined above, may also be produced by reacting the compounds of the formula (V)

S(CH2)nCOOH /C-NHCH 2 (V)

wherein n is the same as defined above, or a reactive derivative

thereof, with a compound of the formula (VI)

R3 OH (VI)

wherein R3' is the same as defined above. •The reaction is carried out by the same procedure of the

above mentioned esterification.

The thus obtained compounds of the formula are then

isolated by conventional techniques.

C Usually, it is best to recover the present compounds in the

form of their acid addition salts as above mentioned. The compounds of the formula may contain crystal water

in equimolar amounts.

The thus obtained compounds of the formula have an excellent

S range of pharmaceutical activity. That is, the present compounds will

"i Z exhibit excellent inhibitory effects on protease, such as trypsin, chymotrypsin, thrombin or urokinase. The present compounds also exhibit excellent anti-ulcer effects. They can thus be used as an excellent preventive or for healing ulcer models in rats such as

Shay ulcers, stress ulcers, indomethacin ulcers, acetic acid-induced

14'i ulcers, cysteamine ulcers or histamine ulcers. 3*

These compounds were found to strongly inhibit the volume of

*the gastric secretion, the acidity of the gastric juices and peptic

activity; Moreover, these effects appear to be quite long lasting.

Experiments of acute and subacute toxicity confirmed that the present

compounds have a low degree of toxicity. Particularly, the compounds

of the formula (II) exhibit excellent anti-ulcer effects and appear

to be remarkably safe.

Compounds of the present invention also exhibit anti-histamine

effects, anti-inflammatory effects and anti-allergic effects (passive

cutaneous anaphylaxis test). The compounds of the formula have excellent inhibitory

effects on protease. The inhibitory effects were determined by the

following methods:

1) Inhibitory Effects on Trypsia:

Inhibitory effects of the compounds on trypsin were determined

according to the method described by M. Muramatsu et al (The Jourrial

of Biochemistry, Volume 58, 214 (1965)). In particular, inhibitGry

effects of the compounds on hydrolysis of p-tosylarginine methyl &9ter

by trypsin were examined (incubated at 370 C for 10 minutes).

2) Inhibitory Effects on Chymotrypsin:

Inhibitory effects on the compounds of chymotrypsin were

determined according to the method described by M. Muramatsu (The

Journal of Biochemistry, Volume 62 408 (1967)). In particular,

inhibitory effects of the compounds on hydrolysis of N-acetyl-L-.

t.yrosine ethyl ester by chymotrypsin were examined (incubated at 0 S37 C for 10 minutes).

3) Inhibitory Effects on Thrombin:

Inhibitory effects of the compounds on thrombin were

determined according to the method described by M. Muramatsu et al

(The Journal of Biochemistry, Volume 65 17 (1969)). In

particular, inhibitory effects of the compounds on hydrolysis of

p-tosy- arginne methyl ester by thrombin were examined (incubated

at 37C for 10 minutes).

4) Inhibitory Effects on Urokinase:

0* Inhibitory effects of the compounds on urokinase were

determined according to the method described by A.J. Joeson et al

(Throm. Diath Haemorrh. 21, 259-272 (1969)). In particular,

inhibitory effects of the compounds on hydrolysis of N-acetylglycyl 0 lysine methyl ester by urokinase were determined (incubated at 37 C

for 10 minutes).

The results obtained are shown in 1. able

FIT;

TABLE 1 INHIBITORY EFFECTS ON PROTEASE

inhibition(% *9 6g. 0 Test Compound Trypsin Chymotrypsin Thrombin Urokinase 660 0 o e~ Compound 1 79 100 37 32 *e

4 Compound 2 61 100 51 72 *d q 9. Clompound 3 42 48 30 52 .o

Compound 4 22 28 35 84 Compound 5 38 90 34

6000 Compound" 6 25 70 39

~t* Compound 7 23 70 32 67 9 #0

Compound 8 51 20 43 ~4 6 *9 .46 Compound 9 28 60 29 Co~ihpound A 65 0 30 32 4 0 ~0 0 Compound 1: 2'-benzyloxycarbonylphenyl tans-4-guanidinomethyl- 9 *09990 0 cyclohexanecarboxylate hydrochloride Compound 2: 2'-methoxy-4' -formylpheny 1 "trans-4-guanidinomethyl- cycl ohexarnecarboxyl ate hydrochlooride Compound 3: phenyl tranis-4-guanidi nomethyl cyci ohexanecarboxylata hydrochloride Compound 4: 2tLethoxyphenyl trans-4-guani di nomethyl cyci ohexane,- carboxyl ate hydrochloride Compound 5: 2Lphenoxycarbonyl phenyl tran5-4-guanidinomethyl cyci ohexanecarboxyl ate hydrochlooride

C h* ~:Y

11

Compound 6: 4'-ethoxycarbonylphenyl trans-4-guanidinomethyl-

cyclohexanecarboxylate hydrochloride

Compound 7: 3'-pyridyl trans-4-guanidinomethylcyclohexane-

carboxylate hydrochloride

Compound 8: 4'-(2"-ethoxycarbonylethyl)phenyl trans-4- 0 4+ guanidinomethlcyclohexanecarboxy'late hydrochloride 4

*r 4 Compound 9: 2'-ethoxycarbonyphenyl trans-4-guanidinomethyl- 00 0* 0* cyclohexanecarboxylate hydrochloride

Compound A: 4'-(2'Lhydroxycar. y ;ethyl)phenyl trans-4-amino-

methylcyclohexanecarboxylate hydrochloride

goe (described in J. Med. Chem., Vol. 15, No. 3, 0r 0000 247 (1972))

1. Shay Ulcers

Male Sprague-Dawley strain rats, weighing 160-180 g were

deprived of food but allowed free access to water for 20 hours prior

to experiment. Under ether anesthesia, the abdomen was incised and

the pylorus was ligated as described by Shay et al (Gastroenterology,

43 (1945)).

The rats were sacrificed 18 hours later by an overdose of ether

and the stomachs removed. The stomach was incised along the greater

curvature and the surface area of each lesion in forestomach was

measured by the naked eye. The lesion was arbitrarily graded into

6 degrees as an ulcer index according to the method of Adami et al

(arch. int. Pharmacodyn., 147, 113-145 (1964)) as follows:

.j jif

0 no lesion 1 hemorrhagic suffusion

2 1-7 small ulcers 3 mm)

3 many small ulcers (more than 5) or 1 ulcer of marked size

4 many ulcers of marked siz.i

4. 5 perfor ted ulcer

The.test drugs were given intraperitoneally, immediately after *0 pylorus ligation. The results obtained are shown in Table 2. 4

TABLE 2

SHAY ULCERS

Test Dose Ulcer Grade *i 0 Compounds (mg/kg) _otal 0 1 2 3 4

Control 0 1 0 2 2 5

Compound 1 300 7 0 0 0 0 0 7

Compound 1 150 10 0 0 0 0 0

Compound 1 75 8 0 2 0 0 0

Compound 3 400 11 4 5 0 0 0

Compound 8 200 4 2 2 1 1 0

Compound B 12.5 2 1 0 1 3 3

Compound A 300 6 0 0 1 2 1

Compound A 150 2 0 3 1 4 0

Compound B: atropine sulfate

Compound 1, Compound 3, Compound 8 and Compound A are the same

as defined above

L

2. Acetic Acid Ulcers

Male Sprague-Dawley strain rats, weighing about 200 g were

used. Under ether anesthesia 10% acetic acid solution (0.05 ml) was

injected carefully between the serous membrane and the muscle near

the pylorus, and the abdomen was closed. Thereafter, the animals

were maintained under conditions and administered 1 ml/100 g of each

test drug dissolved or suspended in 0.5% carboxymethylcellulose

I solution p.o. daily x 10 starting from the next day of the operation.

On the llth day, the rats were sacrificed under ether anesthesia, and the

to stomachs were removed. The area of the ulcer was measured and graded

into 5 degrees as an ulcer index according to the following method: Ulcer Index The Ulcer Area

1 0 10 mm 2 2 11 20 mm 2 3 21 30 mm

4 31 40 mm2 2 5 40 mm

The results obtained are shown in Table 3.

TABLE 3 ACETIC ACID ULCERS

Test Compounds IDose I j(mg/kg) f 1 Ulcer Grade 2 3 4 5 Total *so*

Control Compound 500 Compoundc 250 900 Compound 125 Compound 500 SO0 05 00 a Compound 250 Com~pound 125 Compound 300 Compound 500 Compound 250 Compound 125 Compound 500 Compound 300 Compound 500 Compound 300 Compound 250

Compound 10: 2'-hydroxycarbonyiphenyl trans-4-guianidinomethyl-- cycl ohexanecarboxyl ate hydrochloride Compound 1, Compound 2, Compound 8, Compound 9 and Compound A are the same as defined above.

3. Duodenal Ulcers (CysteatrOne-Induced) Female Sprague-Dawley strain rats, weighing about 200 g

were deprived of food for 24 hours prior to experiment. Cysteamine

400 mg/kg was given subcutaneously once to the rats. The animals

were provided food and libitum from the 7 hours later. The test drug was given orally as aqueous suspension after 7 hours, the a.

administration of cysteamine, then daily x 4. On the next morning

9* of the last administration, the animals were sacrificed under ether I 2 anesthesia. The ulcer area (mm was measured and described as ulcer

index.

The results obtained are shown in Table 4.

TABLE 4

CYSTEAMINE-INDUCED ULCERS a** S @4 *a S

Test Dose Number Ulcer 2 1ndex Inhibition Compounds (mg/kg) of Rats (mm 4

a

*549S* Control 7 80.7 4.3 ~b

Compound 1 500 6 11.7 3.6 85.5

Compound 1 250 6 22.5 3.9 72.1

Compound 1 is the same as defined above.

4. The Influence to the Gastric Secretion

Male Sprague-Dawley strain rats, weighing 160-180 g were

deprived of food but allowed free access to water for 20 hours prior

to experiment. Under ether anesthesion, the abdomen was incised and

the pylorus was ligated as described by Shay et al (Gastroenterology,

43 (1945)). The test drug was given intraperitoneally immediately

after pylorus ligation. The rats were sacrificed 3, 6 and 12 hours

later and the stomachs removed. The gastric juice was collected and

analyzed for volume. Then the gastric juice was centrifuged at 1300 g

for 10 minutes at room temperature to give the supernatant. The total

acidity and peptic activity of the supernatant were determined. Total

acidity was determined by titrating with 0.01 N NaOH. Peptic activity

was determined according to the method described by Anson et al (J.

Gen. Physiol., 22, 79-89 (1938)). The results are shown in Tables 5-7.

(Total (0 acidity is shown as the titration volume (ml) and peptic activity

is shown as the corresponding weight (mg) to crystal pepsin.

O"Sf

I, Sv elm« t

4~ 0 0 0 0@ 0* 0 COO 0 000 3 '4" 0# 00 0 0 0* *0 0 0 0 0 0 0 0S 0 0 0 0 00 C 0 0 00 0 4~ 4. @0 see ~0 000 *~0 *0 0 0 ~j *00 U 80~ 0

TABLE (3 Hours After Trial)

Gastric Contents Test Dose Number ITtatable Acid J Coimpounds (mg/kg) of Rats Volume j output Output (tml) cang 11 gi C e (Img) %hag

Control 5 2.9± 0.2 182.6±+10.6 517.8 0.9

Compound 1 300 5 0.5± 0.2 82.8 14.6± 3.3 94,4 10.9 0.6 84.5 Compound 1 150 5 0.7± 0.3 75.9 9.8± 3.3 88.1 [1.4 0.7 75.9 Compound 1 75 5 2.6± 0.4 10.3 39.2± 11.6 52.5 5.7 1.5 1.7 Compound B 12.5 5 1.2± 0.2 58.6 17.7± 7.7 7-8.6 3.3 0.9 43.1 Compound C 300 5 1.4± 0.3 51.7 22.2± 11.5 73.1 3.1 0,8 46.6 Compound C 150 5 2.1± 0.3 27.6 4.0± 4.0 95.2 j5.6 0.5 3.4 Compound C 75 5 3.2± 0.4 -10.3 44.7± 12.6 45.9 9.6 2.3 -65.5 a SW 0 660 a S «a ae C 6 S S SO 606 a 0 0 0S 50 6 0 5 6 S 6 0S 0 0 8 00 6 9 0 «i a a a. a 005 600 0* 4 e0 Ot. 0 6 S @00 a 66,5 0

TABLE

(3 Hours After Trial)

Gastric Contents Test Dose Number Titratable Acid Titratable Pepsin Compounds (mg/kg) of Rats Volume Output Output

(ml) change (Lg) change )mg) change 4- 4- 6-

Control 2.9± 0.2 82.6 10.6 5.8 0.9

Compound 300 0.5± 0.2 82.8 4.6± 3.3 94.4 0.9 ±0.6 84.5

Compound 150 0.7± 0.3 75.9 9.8± 3.3 88.1 1.4 ±0.7 75.9

Compound 75 2.6± 0.4 10.3 39.2± 11.6 52.5 5.7 ±1.5 1.7

12.5 1.2 0.2 58.6 17.7± 7.7 78.6 3.3 ±0.9 43.1 Compound

Compound 300 1.4± 0.3 51.7 22.2± 11.5 73.1 3.1 ±0.8 46.6

Compound 150 2.1 0.3 27.6 4.0± 4.0 95.2 5.6 ±0.5 3.4

Compound C 75 3.2± 0.4 -10.3 44.7± 12,6 45.9 9.6 2.3 -65.5

8 4 t~4 a. ~4 *ea 60 8 8 b 0 400 4* **o lh 0

TABLE 6 (6 Hour's Afteri Trial)

I IGastric Contents Test Dose Compounds (mg/kg) iNumber of Rats Volume 'Titratable Acid Titratable Pepsin, tput Output

(ml) change (pEq) change NOg change i I i i i I Control 6.1+0 .6 381 .9+67.5 21 .7+3. Compound I 300 1.5+0.3 75.4 70.4+10.4 81.6, 3.5+1.4 83.9 150 1 .5+0.3 75.4 85.3+49.4 7-7.7 4.7+1. I 78.3 4.2+0.6 31 .1 259.8+61.8 32.0 13.9+3.0 315.9 Compound B 12.5 2.7+0.6 55.7 171 .6+50.0 55.1 9.7+1 .6 55.3 Compound C 300 2.1+0.7 65.6 45.3+26.3 88 ,1 8.9+1.7 59.0 150 4.U±0. 5 21 .3 290.6+70.5 23.9 18.7+2.2 13.8 4.3+ 0.5 29.5 220. 3+30. 4 42.3 15.2+1.5 30.0 f 4 I I I

Compound 19 B and C are the same as defined above.

S a Se 55 SSOS e 5B*CO 0 6 0* e 0 5 5 0* S S S S 5 55 @5 Ge S S G:a *6 SOS

TABLE 6

(6 Hours After Trial)

Gastric Contents Test Dose Number Compounds (mg/kg) of Rats Volume Titratable Acid Titratable Pepsin Output Output

(ml) change (pEq) change (mg) change

Control 5 6.1+0.6 381.9+67.5 21.7+3.4

Compound 1 300 5 1.5+0.3 75.4 70.4+30.4 81.6 3.5+1.4 83.9

150 5 1.5+0.3 75.4 85.3+49.4 77.7 4.7+1.1 78.3

5 4.2+0.6 31.1 259.8+61.8 32.0 13.9+3.0 35.9

Compound B 12.5 5 2.7+0.6 55.7 171.6+50.0 55.1 9.7+1.6 55.3

Compound C 300 5 2,1+0.7 65.6 45.3+26.3 88.1 8.9+1.7 59.0

150 5 4.8+0.5 21.3 290.6+70.5 23.9 18.7+2.2 13.8

5 4.3+ 0.5 29.5 220.3+30.4 42.3 15.2+1.5 30.0

Compound 1, B and C are the same as defined above.

cc 0 e c "a 8O S *w C a 0 4 0 0 .c 0 el~ C C C S 0 c cc SO 0 C SC 0 eec cern CO eec eec c cc. a eec

TABLE 7 (12 Hours After Trial

V Test Dose Number Gastric Contents Compounds (mg/kg) of Rats Vol ume Titratalle /Acid. Ii urtbeu epi Outnut output (ml S% change (PEq) change (mg) change -4 -V f -F Control 13.5+1 .8 820.6+100.0 58. 0+9 .8

,Comipound 1 300 3.6+0.6 73.3 258. 4+66 .9 68.5 14.0+2.4 75.9

150 6 .3+0.9 53.3 387.3+79.3 52.8 24.0+4.2 58.6

5.2+2. 1 61 .5 394. 6+1 36 .5 51 .9 27.3- '8.6 52n.9

Compound B 8.1+0.4 40.0 617. 1+48 .0 24.8 34.2+'!.4 41 .0

Czurpound C 300 8,.2+0 e4 39.3 597.0+31.4 27 .2 34.2+21 .9 41.0

150 9.5+0.4 29.6 638.0+22.7 22.3 41 0 29.0

12.0+0.9 11.1 746.6-:- ?.8 9.1 45.9+3.6 20.9

Compound 1, B and C are the same as defined above.

0 *0 @66 S C 00 S 000 *0 C .a S C S C S 0 5 *5 S 0 C S N S Ce C C 0 S C C C 0 000e S C C C 'C 0 9 @00 CC C 6

TABLE 7

(12 Hours After Trial

Test Dose Number Gastric Contents Compounds (mg/kg) of Rats Volume Titratable Acid Titratable Pepsin

Output Output

(mI) change (iaEq) change (mg) change

Control 5 13.5+1.8 820.6+100.0 58.0+9.8

Compound 1 300 5 3,6+0.6 I 73.3 258.4+66.9 68.5 14.0+2.4 75.9

150 5 6.3+0.9 53.3 387.3+79.3 52.8 24.0+4 2 58.6

5 5.2+2.1 61.5 394.6+136.5 51.9 27.3+8.6 52.9

Compound BI 12.5 5 8.1+0.4 40.0 617.1+48.0 24.8 34.2+1.4 41.0

Ccmpound C 300 5 8.2+0.4 39.3 597.0+31.4 27.2 34.2+21.9 41.0

150 5 9.5+0.4 29.6 638.0+22.7 22.3 41.2+2.0 29.0

5 12.0+0.9 11.1 746.6+52.8 9.1 45.9+3.6 20.9

Compound 1, B and C are the same as defined above.

The toxicity of the typical compound of the present compounds is described in the following: 1.Acute Toxicity. Normal ICR strain mice male: 25 .x27gfemale: 22 nu 24g) were used. The test drugs were given orally using gastric sonde.

e~ a Animals were observed for 7 days. The LD50 value was calculated by the

00 S Probit method Bliss). The res-ults obtained -are shown in U. A~0O OS 0 4~ 0 a Table 8. 00 0 "0 9 00 0@ *e 0 TABLE 8

I, Test LOSO (mg/kg) A 0 Compounds Ma Ie male 0000

S Compound 1 >8000 7600 00

0

Compound I is the same as defined above.

e 0 ~4)

I ~A 2. Sub-acute Toxicity p

Q Sprague-Dawley straNin rats, weighing about 150,g were used. The dose was administered onan a day, one-month. Dosages were set at 41,',evels: 40,-,g/kg, 130 mg/kg, 400 mg/kg, 1300 mg/kg of xycarbonylphenyl trans-4-guaiidi nomethyl cycl ohexanecarboxyl ate hydrochloride. The following items were observed. 1) Observations of general symtoms, body weight, food-intake and water-intake. 2) Hematological[ examinations

3) Serum-biochemical examinations

4) Organ weight

Histopathological examinations

As the results, 4 rats died only at the highest dose (1300

mg/kg), but toxic lesions were not observed at the other doses.

Certain compounds of the formula exhibit anti-histamin

effects. For example, 1 5 -g of phenyl trans-4-guanidino- C

methylcycl ohexanecarboxylate hydrocnloride inhibited completely the 6 contraction of the ileum of a guinea pig induced by 10 g/ml

of.histamin dihydrochloride.50 pg of phenyl trans-4-guanidino-

methylcyclohexanecarboxylate hydrochloride inhibited completely the

contraction of the ileum of a guinea pig, sensitized with egg

albumin, when the compound was given with 250 .ig/ml egg albumin

as antigen. Further,- 20wfl/k-eo- phenyl trans-4-guanidinomethyl-

cyclohexanecarboxylate inhibited -akeit 50% carrageenin-induced SE- aL6Jbo 200 my/1r Sinflammation (Wister strain rats, When the compounds of o C

the. present invention are used as anti-ulcer agents, particularly a* 0

prefered are the compounds having the formula These

compounds exhibit both oral and parenteral activities, but, of

L0 course, oral would be the preferable mode of administration. Oral L

administration can be made by capsule, tablet, powder or granule.

In the dosage form, the active compounds are admixed with at least

one inert diluent, such as lactose, corn starch, crystalline

cellulose; a lubricant, such as magnecium stearate; a binder such

as hydroxy propylcellose; a coloring material; perfumery;

(AC/L sweetening agent; or the like.

The dosages of the compounds of this invention in various

compositions actually utilized may be varied. However, it is

necessary that the amount of the compounds be such that two

suitable dosage forms -re attained. Any selected dosage depends *b.9

upon the desired therapeutic effect, administration route and

treatment duration. Such dosage lies, in general, in a range

from 50 1500 mg/day. This invention is illustrated in S** further detail with reference to certain specific Examples, which are presented herein for purposes of illustration only and are

not to be construed as necessarily limiting of the invention. 0

eo o0 00

0

a

Example 1.

Vitamin E trans-4-guanidinomethylcyclohexanecarboxylate hydrochloride: *6*9

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride (11.8g), Vitamin E (17.2g) and dicyclohexylcar-

bodiimide (12.4g) in pyridine (150 ml) was stirred at room

temperature for 30 hours. After removal of insoluble materials

by filtration, the filtrate was evaporated to dryness and the

residual solid was treated with a mixture of 0.1 N-hydrochloric

acid (200m1) and ethylacetate (100 ml) for 1 hour. The *a 1 O insoluble materials were removed. The organic layer was

filtered, concentrated by filtration, and ether also added to

furnish a pale yellow crystal, Vitamin E trans-4-guanidinomethyl-

cyclohexanecarboxylate hydrochloride (16.1 g, m.p. 183- -1 186°C. IR: max(cm-) 1740

Analysis: Found: C, 70.62, H, 10.45, N, 6.29, C3 8 H6 5N3 0 3 .HC1

requires: C, 70.39, H, 10.26, N, 6.48.

S0.1 mM of this compound inhibited about 50% hydrolytic activity by

thrombin and trypsin.

EkxampIe 2.

4'-(2"-Benzyloxycarbonylethyl)phenyl trans-4-guanidinomethylcyclo-

hexanecarboxylate hydrochloride: a6rr A suspension of trans-4-guanidinomethylcyclohexanecarboxylic u* acid hydrochloride (7.19g), benzyl 4-hydroxyphenyipropionate

0# and dicyclohexylcarbodiimide (7.2g) in pyridine (75 ml) was stirred

at 25'C for 15 hours. After removal of insoluble materials by

filtration, the filtrate was concentrated under reduced pressure.

The residue was treated with a mixture of 0.1 N-hydrochloric acid

(100 ml) and ethyacetate (50 ml), the resulting solid was removed U by filtration. The organic layer was filtered, concentrated and r*#9

the residual gummy materials were treated with ether and stirred

to furnish white crystals which on recrystallization from

methanol-ether, gave 4'-(2"-benzyl oxycarbonylethyl )phenyl trans-4-

guanidinomethylcyclohexanecarboxylate hydrochloride (13.lg, 92.1%),

m.p. 77-80 0C.

IR: vmax(cm-j 1745, 1725 (C=0)

Analysis: Found: C, 62.98, H, 6.65, N, 9.04. C2 5 H3 1 N3 04 HC1

requires: C, 63.55, H, 6.80, N, 8.86.

LKarnple 3.

##SIP 4'-(2"1-Ethoxycarbonylethyl )phenyI trans-4-guanidinotmethylcyclo- 4 0~ hexanecarboxyl ate hydrochloride: A suspension of trans-4-guanidinoiethyl cyclohexanecarboxyl ic

acid hydrochloride (11.8 ethyl 4-hydroxyphenylpropionate (10.7 g) and dicyclohexycarbodiimide (11.4 g) in pyridine (150 ml) was stirred at 2?50C for 15 hours. After removal of insoluble materials by

a .::filtration, the filtrate was evaporated. The residue was treated go~ with 1N hydrochloric acid (150 ml), the resulting crystals were removed by filtration and the filtrate was washed with ether. The

aqueous layer was concentrated and the residue was treated with ether to furnish which crystals which on recrystallization from ethanol/ether, gave 4'-(2"'-ethoxycarbonylethyl )phienyl trans-4- aguanlidinomiethylcyclohexanecarboxylate a hydrochloride (17.9 g, 86.9%),

Mn.P. 90-910C.

(cm'1 1725, 1740 (C=O) max Analysis: Found: C, 57.98, H, 7.10, N, 10.13, C20 H29 N304 HC1 requires: C, 58.32, 14,7.34, n, 10.24.

ExDaTple 4. 44 P henyltran5-4-g3-uaniditomthyleyclobiexanecarboxylate

hydrochioride:

4 A suspension of trans-4-guandinomethylcyclohexanecarboxylic

*acid hydrochloride (11.8 phenol (5,6 and dicyclohexyl- *carbodiimide (12.4 g) in pyridine (75 ml) was stirred overnight

at room temperature. After evaporation of solvent, the residue

was treated with 0.1 N hydrochloric acid (200 ml), the

**insoluble materials removed by filtration and the filtrate was

washed with ethylacetate. The aqueous layer was concentrated to 100 ml, the resulting crystals were filtered and washed

with isopropylalcohol/isopropylether to give phenyl trans-4-

guanidinomnethylcyclohexanecarboxylace hydrochloride (12.5 g, r m.p. 150-153,C. rhis compound was recrystallized from

methanol to give white crystal, phenyl trans-4-guanidinomethyl-

cyclohexanecarboxylate hydrochloride, m.p. 159.5-161.5 0 C.

ma.R,: 1 1750 1620-1680 max (cm Analysis: Found: C, 57.49, H, 7.25, N, 13.27, C15 H2 1N3 0 2.HCl1

requires: c, 57.78, H, 7.11, N, 13.48.

Ex;aple

4'-(2"'-Carboxyethyl)phenyl trans-4-guanidinomethylcyclohexane-

carboxylate hydrochloride:

A suspension of trans-4-guanidinomethyicyclohexanecarboxylic 0*

acid hydrochloride (11.8 benzyl 4-hydroxyphenylpropionate e4 4

4.4 (15.4 g) and dicyclohexylcarbodiimide (14.4 g) in pyridine (80 ml)

was stirred overnight at room temparature. After evaporation of

solvent, the residue was treated with a mixture of 0.1 N-hydro-

chloric acid (200 ml) and ethylacetate (100 ml), the insoluble

0 104 materials were filtered off and the organic layer was separated. 04

After evaporation to dryness methanol, acetic acid and water were.. added, then, the resulting clear solution was hydrogenated over

Pd/C. After absorption of theoretical amount of hydrogen,

the catalyst was filtered off. The filtrate was concentrated

to dryness, the crystals obtained were recrystallized from

methanol/acetic acid to give the title compound (14.3 g, 74.5%),

m.p. 295-296°C.

vmax(cm 1750(C=0), 1706(c=0), 1630-1680(C=N)

Analysis: Found: C, 55.98, H, 6.51, N, 10.72.C 18 H2 5N3 04HCI

requires: C, 56.32, H, 6.83, N, 10.95.

Example 6.

2'-Benzyloxycarbonylphenyi trans-4-guanidinomethylcyclohexanecarboxylate

hydrochloride: 4A suspension of trans-4-guandinomethylb.-yclohexanecarboxylic acid hydrochloride (10.6 benzyl salicylate (11.4 g) and dicyclohexyl- carbodilmide (11.3 g) in pyridine (100 ml) was stirred at 35-40 0C

for 15 hours. After removal of insoluble materials, the-solvent was eyaporated. The residue was treated with 0.1 N-hydrochloric acid (200 ml), the resulting crystals were obtained, and extracted

with methanol. The extract was concentrated and water was added, the resulting solid was recrystallized from aqueous acetone to givye 2'-benzyl oxycarbonyl phenyl trans-4-guanidi narethylcyclohexanecarboxyVlate hydrochloride (17.9 g, m.p. 70-72.5*C. v max (Cm-1 1730 (C=O) AnAlysis: Found: C, 61.38, H, 6.38, N, 9.19, C23 H27 N3 04 HC1

requires: C, 61.95, H, 6.33, N, 9.42 The product obtained was recrystallized from methanol/ether to give a white crystal, 2'-benzyloxycarbonylphenyl trans-4*-guanidinomethyl- cyclohexanecarboxylate hydrochloride, m.p. 83'C.

-29.-

Exanle 71

2'-Hydroxycarbonylphenyl trans-4-guanidinomethylcyclohexanecarboxylate 0* hydrochlooride:

4 A solution of 2'-benzyloxycarbonylphenyl trans-4-guanidino- rnethylcyclohexanecarboxylate hydrochloride.,(8 .9 g) in acetic acid (30 ml) and methanol (10 in!) was hydrogenated ovc"' 10% Pd/C. After absorption of hydrogen (about 500 ml), the catalyst was filtered off. The filtrate was concentrated to dryness, the crystals *000 were washed with ether, and recrystallized from ethanol/ether to give the title compound (6.5 g, m.p. 166-168'C.

vma (cnf1 1750, 1690(C=G) Analysis: Found: C, 53.85, H, 6.05, N, 11.42, C16H21N304 'HC requires: C, 54.01, H, 6.23, N, 11.81.

Example 8.

2'-Ethoxycarbonylphenyl trans-4-guanidinomethylcyclohexanecarboxylate CQ p 008 hydrochloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic

acid hydrochloride (7.1 ethyl salicylate (5.5 g) and dicycio- *o0

S hexylcarbodiimide (7.2 g) in pyridine (100 ml) was stirred at room

temperature for 15 hours. After removal of insoluble materials by

filtration the solution was concentrated. The residue was treated

with 0.1 N-hydrochloric acid (100 ml), further insoluble materials 0* "X1. were filtered off and the filtrate was washed with ether. After

evaporation, the residue was treated with ether, and stirred, the

resulting crude product was recrystallized from ethanol/ether

to give 2'-ethoxycarbonylphenyl trans-4-guanidinomethylcyclohexane-

carboxylate hydrochloride (8.4 g, m.p. 110-l111C. 1 vmax (cm 1740 (C=0)

Analysis: Found: C, 55.97, H, 6.72, N, 10.54. C1 7 H2 5 N3 0 4 .HC1

requires: C, 56.32, H, 6.83, N, 10.95.

Exapnle 9.

2'-Methoxy-ad' -rormylphenyl trans-4-guanidinomethyl cyc lohexane-

carboxylate hydrochloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic

acid hydrochloride vaniline (5.0 g) and dicyclohexyl-

carbodiimide (7.2 g) in pyridine (100 ml) was stirred at 300 C

for 15 hours. After removal of the insoluble materials the

solution was concentrated. The residue was treated with a Ouse rixture of 0.1 N-hycrochloric acid (100 ml) and ethylacetate **U

O (100 ml), and stirred for 1 hour. Further insoluble materials pe

were filtered off and the organic layer was separated. After ethger awuxo 1-O-n ,6oPYohD/e~lie~ ±D ZDY evaporation to dryness, the residual solid was washed with(

2'-methoxy-4'-formylphenyl trans-4-guanidinomthylcyclohexanecarboxylate hydrochloride (9.6 g, m.p. 110-1110 C.

V 1760 (C=0) max Analysis: Found: C, 54.74, H, 6.66, N, 11.22. C1 7H2 3 N304 HC1

requires: C, 55.21, H, 6.54, N, 11.36.

4U,.

Mcarnple

2'-Phenoxycarbonyiphenyl trans-4-guanidinomethylcyclohexane-- carbodyl ate hydrochloride: suspension of trans-4-guanidinomethylcyclohexane carboxylic hydrochlor"ide (27.5 phenylsalicylate (25.0 g) and 0 dicyclohexylcarbodiimide (26.5 g) in dimethvlformamide (100 ml)

was stirred at roomi temperature for 21 hours. To this solution, water (150 ml) and concentrated hydrochloric acid (120 ml) were

OOSS added, the resulting precipitated solid was washed with water. Then, the solid was treated with methanol and the methanol layer was

evaporated to dryness and solidified with ether to give 2'-phenoxy.- carbonyl phenyl trans-4-guanidi nomethyl cycl ohexanecarboxyl ate hydrochloride (34.1 g, m.p. 157-162'C. vmx(cm 1 1740, 1750 (C=0) **goo NI4R: 6CD 3OD

0.7-3.1 (in, 12H) 6.8-8.1 (in, 9H)

Analysis: Found; C, 61.08, H, 5.95, N, 9.79. C22H25 N3 04 HC1 requires: C, 61.18, H, 6.07, N, 9.73.

Example 11.

3' -Berzyloxycarbolnylphenyl trans-4-guanidinoaethylcyclohexanecarboxylate

hydrochloride:

**3'-Benzyoxycarbonylphenyl trans-4-guanidinamethylcyclohexane-

carboxylic acid hydrochloride (109.8 benzyl 3-hydroxybenzoate

(106.3 g) and dicyclohexylcarbodiimide (105.7 g) in pyridine (450 mi) 4. was stirred at room temperature for 22 hours. After evaporation of a.* a 0 pyridine, water (100 ml) was added and acidified with hydrochloric

acid. The resulting slurry was treated by centrifug. separator. (1 The solid obtained was treated with merthanol, and the methanol layer

was concentrated. Recrystallization of the residue from isopropyl-

alcohol gave 3'-benzyloxycarbonylphenyl trans-4-guanidinomethyl-

C' cyclohexanecarboxylate hydrochloride (169.5 g, 81.6%) m.p. 75-80 0 C.

max(cm- 1 1725, 1755 (C=0) max NMR: 6CD3 0D

0.8-3.2 12H)

5.3 2H)

6.9-8.0 9H)

Analysis: Found: C, 61.37, H, 6.18, N, 9.58. C23 H27 N304 HCl

requires: C, 61.95, H, 6.33, N, 9.42.

Example 12. 3 '-Hydroxycarbonyl tranL---4-guanidionrethyl~ccohexanecarboxylate hydrochloride:

0* 3'-Benzyloxycarbonylphenyl trans-4-guanid-inomethyl cyclo- *6 hexanecarboxylate hydrochloride (80 g) was dissolved in a

ixture of methanol (300 ml) and acetic acid (300 ml), and the solution was hydrogenated over 10% Pd/C. A-fter absorption of theoretical amount of hydrogen, the catalyst was filtered off. The filtrate was concentrated to dryness, the crystals

IV 00 obtained were recrystallized from mnethanol to give the title so:. zoirpound (56.8 g, m.p. 197-200 0 C. I.R:vmax(cm 1 1700, 1740 (C=0) NMR: 6CO 3

0.8-3.2 (mn, 12H) 7.0-8.0 (m,4H) Analysis: Found: C, 53.96, H, 6.21, N, 11.89. C16 H21 N3 04 HCl requires: C, 54.01, H, 6.23, N, 11.81.

Example 13.

4'-Ethoxycarbonyl phenyl trans.-4--guani di nomethyl cyci ohexane- carboxyl ate hydrochlooride: A suspension of trans-4-guanidinomethylcyclohexanecarbo- a0 xylic acid hydrochloride(35.4g), ethyl 4-hydroxybenzoate(25g)

and dicyclohexylcarbodiimide(34.0g) in pyridine(35011) was stirred

at room temperature for 17 hours. After evaporation of pyridine, *e *to the residue was added water(300ni), and the mixture was acidified with hydrochloric acid. The resulting white solid was dissolved in LO methanol, and insoluble materials were filtered off. The methanol layer was concentrated and recrystallizati-on of the residue from ethanol gave 4'-ethoxycarbonylphenyl trans-4- guanidinomethylcyclo-

hexanecarboxylate hydrochloride(33.7g, m.p. 181-1840 c. 04 I vmax (cm 1715, 1755(C=0) NMR: 6CD 3OD

0.6-3.1 4.3 (q,2H)

7.1, 8.0(d,d,4H) Analysis: Found: C, 56.21, H, 6.79, N, 11.03. C18 H25 N3 04 *HC1

requires: C, 56.32, H, 6.83, N, 10.95.

h~cample 14.

I Ss 4'-Hydroxycarbonylphenyl tranis-4-guanidinomethy Icyclohexane- carboxylate hydrochloride: By the procedure of Example 12, using 4'-benzyloxycarbonyl- phenyl trans 4-guanidinomethylcyclohexanecarboxylate hydrochloride

00 (65g) as starting material, 4'-hydroxycarbonylphenyl trans-4- guaniidinomethy'icyclohexanecarboxylate hydrochloride(45.8g, 88.3%) S06

w~s obtained. m.p. 225.5-228.00 C. max (cm 1) 1750(C=0) NMR: as0MS0-d 6 020 0.8-3.2 (in, 12H) 7.2, 8.0 4H-) Analysis: Found: C, 53.89, H, 6.21, N, 11.97. C16H21N3 04 HCI requires: C, 54.01, H, 6.23, N, 11.81.

Example 1. 3'-Methoxycarbonylphenyl trans-4-guanidinomethylcyciohexane- carboxyl ate hydrochloride: A suspension of trans-4-guanidinomethylcyclohexanecarboyylic acid hydrochloride(35.4g), methyl 3-hydroxybenzoate22.8g) and qicyclohexylcarbodiimide(34.0g) in pyridine(300m1) was stirred at

room t-emperature for 24 hours. Following removal of insoluble

0000materials and evaporation of pyridine, the residue was acidified 4 with concentrated hydrochloric acid and extracted with chloroformV. VD~ After concentration of chloroform layer, the residual solid was 000,

10 recrystallized from acetone to give 3'-methoxycarbonylphenyl trans- 4-guanidinomethylcyclohexanecarboxylate hydrochloride(20.9g, 37.7%), *1 m.p. 138-147 C. vmx(cm j 1705, 1745(G0O)

NMR: 6CD 3QD 0.8-3.2 (in, 12H) 3.9 3H)

Analysis: Found: 54.93, H, 6.48, N, 11.43. C17H23 N3 04 *HU

0requires: C, 5.5.21, H, 6.51. N, 11.36.

Exanie 16. '9600. 3'-Pyridyl trans-4-guanidinomethyicyclohexanecarboxylate 0.* di hydrochloride: 0*

A suspension of trans-4-guanidinomethylcylohexanecarboxylic .:acid hydrochloride(47,lg), 3-hydroxypyridine(19.0g) and dicyclo- hexylcarbodiirnide(45.4g) in pyridine(40iil) was stirred at room temperature for 24 hours. The solid collected was extracted with methanol, and methanol layer was evaporatedi to dryness. The res-idue was recrystallized from methanol to give 3'-pyridyl trans- 4-guanidinomethylcyclohiexanecarboxylate dihydrochloride(36.1g,

m.p. 180-185 0 C. vax (cnC 1) 1750(C=0) NMR: SCD OD 0.7-3.1 (in, 12H) 7.6-8.6 (mn, 3H) Analysis: Found: C, 47.98, H, 6.24, N, 16.31. C14H20 N402 2HG1 requires: C, 48.15, H, 6.35, N, 16.04.

Eiainple 17. O'Naphthyl trans-4-guanidinomethylcyclohexanecarboxylate hydro-

chloride:

A suspension of trans-4-guanidinome-thylcyclohexanecarboxylic acid hydrochloride(47.1g), -naphtho1 (28.8g) and dicyclohexyl- carbodiimide(45.4g) in pyridine(400ml) was stirrea at room tempera- ture for 24 hours. After evaporation of solvent, to the residue

'POO***was added water(500ml), and the mixture was acidified with hydro- Se. chloric acid. The resulting white crystals were dissolved in

000 mr,rrcnol(500m1). After evaporation of methanol, the residue was crys talIi zed from methanol to give -naphty1 trans-4-guanid~xho-

1 thylcyclohexanecarboxylate hydrochloride(49.lg, m.p. 195-202 0C.

vmx(cm-I 1750(C0O) NMR: 6DMSO-d 6 0.8-3.1 (mn, 12H) 6.9-8 .2 (mn, 7H) Analysis: Found: C, 62.52, H, 6.59, N, 11.89. C1 9H23 N302 HC1 requires: C, 63.06, H, 6,68, N, 11.61.

Example 18.

a-Naphthyl trans-4-guani di nomethyl cycl ohexanecarboxyl ate hydro-

*0 chloride: By the~procedure of Example 17, using a-naphthol(28.8g) instead of a-naphthol as starting material, a-naphthyl trans-4- gu~anidinomethylcyclohexanecarboxylate hydrochloride(36.9g, 51.0%) a was obtained. m.p. 19 1 2 03 0 C

I.R: vax(cm-1 1745(C0O) .NMR: 6C 3 OD

0.9-3.0 (i,12H) 7.2-8.1 (i,7H)

Analysis: Found: C, 62.71, H, 6.62, N, 11.83. C19H23 N302 'HC1 requires: C, 63.06, H, 6.68, N, 11.61.

-41l-

Exanpie 19.

*e00 2-Formyl phenyl trans-4-guani di nomethyl cyci ohexanecarboxylate *0 fj hydrochloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride(94.3g), salicyl aldehyde(50g) and dicyclohexyl- in pyridine(6O0rfl) was stirred at room tempera-

ture for 16 hours. After evaporation of pyridine, to the residue

was added water(2O0ml), the mixture was acidified with hydro- 'a chloric acid. The resulting white solid was filtered and extracted tO with methanol(5O0ml). The insoluble matter was removed by filtra- V. tion and the filtrate was concentrated. To the residue was added acetone(200ml), insoluble materials were removed by filtration. The filtrate was concentrated. The residue was crystallized from

water to give 2'-formylphenyl trans-4-guaniidinomethylcyclchexane- carboxylate hydrochloride(53.2g, m.p. 135 13 8O0C.

I.R:vax(cm1 J 1740(C0O) NMR: 6CD 30D (in, 12H) 5A4 1H)

6.8 (in, 4H)

Analysis: Found: C, 55.98, H, 6.31, N, 12.63. C16H21 N3 03 HC1 requires: C, 56.55, H, 6.53, N, 12.37. 0.1mM of this compound inhibited 56% hydrolytic activity by urokinase.

Example 2'-Methoxyphenyl trans-4-guanidinomethylcyclohexanecarboxylate hydrochloride: A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride(47.1g), o-methoxyphenol (24.8g) and dicyclo- hexylcarbodiimide(45.4g) was stirred at room temperature for 24 to hours. After removal of insoluble materials, the solutio6 was concentrated to dryness. To the residue was added water(300m1), the solution was acidified with hydrochloric a&2id. The resulting

crystals were filtered and recrystallized from isopropylalcohol to a..0 give the title compound(57.5g, m.p. 141-145O C.

I.R:vax(crif1 1760(C=0) *ase: NMR: 6sCD OD a

0.7-3.0 (in, 12H)

3.8 3H)

6.8-7.3 (mn, 4H)

Analysis: Found: C,56.18, H, 7.01, N, 12.31. C16 H26N303 HC1

requires: C, 56.22, H, 7.08, N, 12.29.

0.1mM of this compound inhibited 50% hydrolytic activity by urokinase.

Example 21.

4'-Methoxyphenyl trans-4-guanidinomethylcyclohexanecarboxylate g* hydrochloride:

S A suspension of trans guanidinomethylcyclohexanecarboxylic 0*

acid hydrochloride(47g), p-methoxyphenol(25g) and dicyclohexyl- in dimethylformamide(200ml) was stirred at room

(0 temperature for 23 hours. To the reaction mixture were added

water(200ml), ice(lOOg) and concentrated hydrochloric acid(200ml).

The resulting crystals were filtered and washed with water, and S**O

dissolved in methanol(300ml). The insoluble materials were filtered

off and the filtrate was concentrated to dryness. The residue was

crystallized from methanol to give the title compound (29.2g, 42.4%).

Sm..p. 203-2050 C.

IR.: mx(cm- 1 1745(C=0) vmax NMR: 6CD 3 00

0.-7-3.1 12H)

3;8 3H)

4H)

Analysis: Found: C, 56.19, H, 7.01, N, 12,.35. C16H23 N3 02 'HU

requires: C, 56.22, H, 7.08, N, 12.29.

0.1mM of this compound inhibited 52% hydrolytic activity by urokinase.

Example 22. 2 '-EthoxyPhenYltrans.-4-guanidinomethylcyclohexanecarboxyl ate .:hydrochl ori de:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic

acid hydmochloride(70.7g), o-ethoxyphenol(41.5g) and dicyclo- hexylcarbodiimide( 68.1g) in direthylformamide(300ml) was stirred at to room temperature for 24 hours. To the reaction mixture were added water(100m1) and ccncentrated hydrochloric acid(3S0MIi). The resulting crystals were flltered and washed with water, dissolved in methanol(3"'Onl). The insoluble materials were filtered off and the filtrate was concentrated to dryness. The residue was dissolved

in acetone and treated with ether to give the title compound (65.0g, m.p. 14 4 148 C.

I.R. vm ax (cm-1 1750(C=0) NMR: 6C0.3.OD 0.7,.3.0 (in, t, 2H) 6.7-7.2 (mn, 4H)

Analysis: 'Found: C, 56.94, H, 7.01, N. 12.11. C17 H25 N3 03'HC1 requires: C, 56.38, H, 7.36, N, 11.81.

Example 23.

21-Acetylphenyl trans-4-guanidomethycylohexarecarboxy late I*0*.hyc~rochl on de:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride(34.7g), o-hydroxyacetophenone(20.Og) and S. dicyclohexylcarbodiirnide(33.3g) in pyridine(300m1) was stirred at room temperature for 24 hours. After removal of insoluble

U0 materials, the solution was concentrated to dryness. To the

residue was added water(200m1) and the mixture was acidified with hydrochloric acid, then extracted with chloroform. The chloroform

layer was concentrated to drYniess, and the residue was recrystal- lized from isopropylalcohol to give the title compound(22.7g, m.p. 159-166 C. (cm- 1 7o~ vmax aNMR: SCD 3OD

0.8-3.1

6.9-8.0 (in, 4H)

Analysis: Found: C, 57.47, H, 6.78, N, 12.03. C17 H23 N3 03 'HC1 requires: C, 57.70, H, 6.84, N, 11.88. 0.05mM of chis compound inhibited 50% hydrolytic activity by chymotrypsi

Example 24.

.0.0:4'-Acetylphenyl trans-4-guan-idinomethylcyclohexanecarboxylate

hydrochloride: A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride(35.4g), p-hydroxyacetophenone(20.4g) and tO dicycilohexylcarbodiimide(34.Og) in pyridine(300m1) was stirred at

room temperature for 24 hours. After removal of insoluble @0 materials, the solution was concentrated to dryness. To the

residue was added water(300m1) and the mixture was acidified with hydrochloric acid. The resulting crystals were filtered and recrystallized from etYhanol to give the title compound(42.4g, M.P. 175-180 0C.

(cm) 1750(C=O) vmax% NMR: 6CD 3 00 0.7-3.1 E,0 7.2, 8.0 4H)

I I

-47- Analysis: Found: C, 57.63, H, 6.81, N, 11.92. C17H23N3 03 'HC1 requires: C, 57.70, H, 6.84, N, 11.88.

Example 2'-Phenylphenyl trans-4-gtianidinomethlylcyclohexanecarboxylate 0000

hydrochloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic

acid. hydrochloride(35.4g), o-phenylphenol(25.5g) dicyclohexyl- carbodiimide(34.0g) in pyridine(30knl) was stirred at room temperature for 24 hours.- The solution was concentrated to dryness. to To the residue was added water(300m1), and the solution was

acidified with hydrochloric acid, then extracted with chloroform. 00 The chloroform layer was washed with water and evaporated to give

em:. the title compound(55.5g, m.p. 78-850 C. I.R:vax(cmf') 1750(C=0) NMR: sCD OD' *0.7-3.0 (in, 12H-) 0 6.8-7.5 9H)

Analysis: Found: C, 64.69, H, 6.49, N, 11.04. C21H25 N3 02 *HC1

requifes: C, 65.02, H, 6.76, N, 10.83.

Example 26. flee. C 4'-Phenylphenyl trans-4-guanidinomethvlcyclohexanecarboxylate G0 0 S C 00 04~ Ce hydrochlori de: @e C a CS C A suspension of trans-4-guanidinomethylcyclohexanecarboxylic SO U CC 0* C acid hydrochloride(47.1g), p-phenylphenol(34.Og) and dicyclohexyl- car:bodiimide(44.5g) in dimethylformamide (250ml) was stirred at roo,ni temperature for 24 hours. After removal of insoluble materials, Ce.. To the residue was added C the solution was evaporated to dryness. CC..

@0 e~. water (300m), and the solution was acidified with hydrochloric to acid. The resulting crystals obtained were recrystallized from *eee *0 *0 C

methe'nol to give the title compornd(47.9g, m.p. 185-196O0 C. I.R: ma(cnf- 1 1750(C=rv) C CSCCSS S

C NMR: 6CD 3OD 6906CC 0' 0.8-3.1 12H) 6.9-7.8 9H)

Analysis: Found: C, 64.94, H, 6.57, N, 11.03. C21H25 N3 03 HCI

requires: C, 65.02, H, 6.76, N, 10.83.

gee. Exanple 27. 9. S o 4'-Phenoxycarbonylphenyl trans-4-guanidinomethyicyclohexanecarbaxylate 0r hydrochloride: e* A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid

hydrochloride (22.0 phenyl 4- hydroxybenzoate (20.0 g) and

*r S dicyclohexylcarbodiimide (22.9 g) in pyridine (100 ml) was stirred

at room temperature for 30 hours. After evaporation of pyridine,

5* to the residue was added water (300 ml), and the solution was acidified 0*0*

oo with hydrochloric acid. The resulting solid obtained was extracted

with methanol (500 ml). After concentration of the extract, the residue

was recrystallized fran ethanol to give 4' -phenoxycarbonylphenyl

trans-4-guanidinomethylcyclohexanecarboxylate hydrochloride

(21.5 g, m.p. 166-170 0 C.

I.R.: v (cmn) 1740, 1745 (C=0)

NMR: 6

0.7 3.1 12H)

8.3 9H)

Analysis: Found: C, 61.09, H, 6.12, N, 9.78. C2 2H2 5 N3 04 HCI

requires: C, 61.18, H, 6.07, N, 9.73.

Exaiple 28.

o 3'-Anisyloxyc~irbonylphenyl trans-4-guanidinometLhylcyclohiexane-

carboxylate hydrochloride: A suspension of 3'-carboxyphenyl trans-4-guanidinomethyl- cy'clobexanecarboxylate hydrochloride(7.12g), p-anisalcohol(2.76g) and dicyclohexylcarbodiimide(5.16g) in pyridine(30mi) was stirred at room temperature for 18 hours. To this reaction mixture was

*see added water(lON1i) and the solution was acidified with hydro- *go*

go, chioric acid and stirred for 1 hour. The resulting solid obtained a otowas extracted with methanol(5Oml). After evaporation of the

methanol, the residue was recrystallized from aqueous methanol to

give 3'-anisyioxycarbonylphenyl trans-4--guanidinomethylcyclo- 0:.:hexanecarboxylate hydrochloride(8.4g, m.p. 90-93 0 C.

1'R:vmax 1725, 1755(C=0)

NMR: 6CD 3ODJ 0.8-3.2 12H) 3.8 3H) 5.3 2H)

6.9-8.0 (in, 8H)

2P Analysis: Found: C, 60.35, H, 6.31, N, 8.89. C24 H29N3 05 HC1 requires: C, 60.56, H, 6.35, N, 8.83

J,

-51-

Eample 29. 3'-Hydroxycarbonylphenyl trans-4--guanidinomethylcyclohexane- 0, carboxyl ate hydrochl oride: 41 00 4, By the procedure of Example 12, using 3'-anisyloxycarbonyl-

e0* phenyl trans-4-guanidinomethylcyclohexanecarboxylate hydrochloride as the starting material, the title compound(l.5g, 80,3%10) was obtained.

Eample 30.

4'-Benzyloyycarbonylphenyl trans-4-guanidinomethylcyclohexane- U. 1,carboxylate hydrochloride:

A suspension of trans-4-guaniclinomethylcyclohexanecarboxylic O S acid hydrochloride(119.4g), benzyl 4-hydroxybenzoate(115.6g) and

dicyclohexylcarbodiimide(114.9g) in dimethylformnamide(430m7) was stirred at room temperature for 20 hours. To this reaction mixture was added water (1500m]), and the solution was acidified with hydrochloric acid(500,m]). The resulting solid obtained was ex- tracted with methanol(500m1). After removal of insoluble materials by filtration, the filtrate was concentrated and the residue was recrystallized from methanol/water to give 4'- benzyl oxycarbonyl phenyl trans-4-guani di nomethyl cyci ohexanecarb- oxylate hydrochloride(148.5g, m.p. 13 4 1 38 0 C.

VMaX (cm 1710, 1750(C-0) NMR: 6CD OD

(Y.8-3.2 12H)

0 a 5.,35 2H) a a. 7.2, 8.1 4H)

A7,.4

Analysis: Found: C, 61.47, H, 6.18, N, 9.53. C2 3H 27 N304 HC1

requires: C, 61.95, H, 6.33, N, 9.42.

Go** a a moo* xample 31.

Bey2zlxycarbnY1PhenY1 -crans-4-guanidinanethylcyclohexane- aes

carboxylate hydrochloride:

A suspension of 4'-hydroxycarbonyphenyl trans-4-guanidino-

methylcyclohexanecarboxylate hydrochloride(2.15g), benzylalcohol

(0.659) and 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydro- chloride(.39g) in pyridine(l1ml) was stirred at room temperature

for 30 hours. To this reactio: mixture was added water(50ml), and

the solution was acidified with hydrochloric acid, the resulting

53-.

crystals were filtered and recrystallized from methanol/water to

give the title compound(1.48g, 55.3%).

Example 32. 2'-Benzyloxycarbonylphenyl trans-4-guanidinomethylcyclohexane- carboxy',ate hydrochloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride(5.89g), benzyl salicylate(5.71g) and 1-ethyl-3- (3"-dimethylaminopropyl)carbodiimide hydrochloride(5.75g) in

pyridine(25m1) was stirred at room temperature for 24 hours. To this reaction mixture was added water(5C0nl), and the solution was acidified with hydrochloric acid. The resulting solid was see*

fil'ered and washed with water, and recrystallized from methanol/ ether to give the title compound(9,27g, 83.2%),

Example 33.

3'-Methoxyphenyl trans-4-guanidinomethylcyclohexanecarboxylate hydrochloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride(35.4g), m-methoxyphenol1(18.6g) and dicyclo- hexylcarbodiimide(34.Og) in pyridine(300ml) was stirred at room temperature for 24 hours. After rEtoval of insoluble materials, the residue was acidified with hydrochloric acid and extracted with chloroform. Following concentration of chloroform layer under

:reduced pressure, the residue was washed with water. The resulting solid was filtered and recrystal lized from ethanol to give the title *,*~compound(29.4g, m.p. 125.5-131.5 0 C. cm 1) 1740(C=0) viflVax' NMR: 6CD 30D 1.0-3.1 (in, 12H) see*

00* 3.8. 3H-) 6.6-7.45 4H)

Exampl e 34.

41-Farmylphenyl trans-4-guanidinoinethiylcyclohexanecarboxyl ate hydrochloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride(35.4g), p-hydroxybenzaldehyde(18.3g) and dicyclohexylcarbodiiinide(34.0g) in pyridine(300n1) was stirred at room temperature for 24 hours. After evaporation of pyridine, to the residue was added water(lO0ml), and the solution was acidified with hydrochloric acid. The resulting solid was filtered ano extracted with methanol. The extract was concentrated to dryness and the residue was recrystallized from methanoi to give the title compound(22.5g, m.p. 157.5-163.50 C.

I, max 1750(C=o)

NMR: 6CD 3 0D

S 0.9-3.1 12H)

7.25, 7.95(d,d,4H)

Example

o, 4'-Propionylphenyl trans-4-guanidinomethylcyclohexanecarboxylate

hydrochloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride(35.4g), p-hydroxypropiophenon(22.5g) and

dicyclohexylcarbodiimide(34.0g) in pyridine(300ml) was stirred 0e*#

at room temperature for 24 hours. The resulting crystals were S

filtered and extracted with methanol(500ml). The extract was

evaporated to dryness and the residue was recrystallized from

ZO methanol to give the title compound(39.6g, m.p. 179-185 C.

I.R: ax(cm 1750(C=0) NMR: 6CD 3OD 0.9-3.0 (in, 17H) 7.1, 7.95 4H)

Example 36. os.

:4'-Diphenyl trans-4-guanidinomethylcyclohexanecarboxylate methanesul fonate: By the procedure of Example 26, using trans-4-guanidino- methylcyclohexanecarboxyliic acid methanesulfonate(2.95g) instead (P of-trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochloride,

the title compound(3.2g, 71.4%) was obtained. m.p. 20 7 21 0O0C.

o ccExample 3 7.

0 2'-Benzyloxycarbonylphenyl trans-4-guanidinomethylcyclohexane-

carboxylate p-toluenesulfonate:

By the procedure of Example 6, using trans-4-guanidino- methylcyclohexanecarboxylic acid p-toluenesulfonate(3.71g) instead of trans-4-guanidinomethylcyclohexanecarboxylic acid

hydrochloride, the title compound(4.3g, 73.9%) was obtained.

ma (cm- 1 1735, 1760(C=O) NMR: SC 3 OD

0.7-3.0

5.3 2H) (in, 13H)

Examiple 38. s* $S

1-Naphthyl trans-4-guanidinomethylcyclohexanecarboxylate hydro- chloride:

A suspension of trans-4-guanidinomethylcyclohexanecarboxylic acid hydrochlof-ld6 a-naphtol(1.44g), sulfuric acid(0.05g) and boric acid(0.03g) in a mixture of d'lmgthylsulfoxide(10m1) and was heated under reflux conditions for 20 hours. The resulting water was removed by azeotropy with xylene. Following concentration of the reaction mixture, the residue was chromato- zo graphed on a column of silica gel with chloroform/methanol as eluent

to give the title compound.

Exanule 39.

4'-IDiphenyl trans-4-guitnidinomethylcyclohexanecarboxylate 000B hydrochloride:

A mixture of trans-4-guani di nomethyl cycl ohexanecarboxyl ic :acid hydrochloride(23.Gg), 4-hydroxybiphenyi(17.Og) and phos- ***phorousoxychloride(7.7g) was stirred at 80-85O0 C for 2 hours.

After adding of toluene(5Oml), the mixture was stirred at 8 0 85 0C for further 2 hours. The solvent was removed by decantation and water was added. The solution was set aside overnight in a refrigerator, the resulting white crystals were recrystallized from

methanol to give the title compound(12.8g, 631.3%).

Example

4'-(2"-Benzyloxycarbonylethyl )phenyl trans-4-guanidinomethylcyclo-

hexanecarboxyl ate hydrochloride: A suspsension of 1-carboxyethyl)phenyl trans-4-guanidino- methylcyclohexanecaboxylate hydrochloride(l.92g) in thionylchloride was stirred at 60°C for 1 hour. Following evaporation of excess thionylchloride, the residue was dissolved in chloroform

The resulting pale yellow solution was added at room

temperature to a solution of benzyl.alcohol(0.65g) and triethyla-

mine(0.51g) in chloroform(5ml). After stirring at 35-400 C for

hours, the solvent was evaporated and the residue was solidified

with water to give the title compound(1.4g, 59.1%).

Example 41. a

4'-(2"-Benzyloxycarbonylethyl)phenyl trans-4-guanidinomethyl- Sact cyclohexanecarboxylate hydrochloride:

A suspension of 4'-(2"-carboxyethyl)phenyl trans-4-guanidino-

methylcyclohexanecarboxylate hydrochloride(1.92g) and benzyl alcohol

(5ml) was stirred at 130-1350 C for 10 hours. After evaporation of

excess benzylalcohol, the residue was recrystallized from methanol/

Sether to give the title compound(l.2g, 50.6%).

Excample 42 3 '-Methoxycarbonyl pheny I trans-4-guani di nc'methyl cyci ohexane- carboxylate hydrochloride: A suspension of 3 '-carboxyphenyl tranis-4-guani di nomethyl cyci o- hexanecarboxylate hydrochloride(3.56g) in thionyl chloride(lOml)

.**was refluxed for 30 minutes. Following concentration of the reaction mi.xture, the residue was dissolved in chloroform(lOml), and to the solution was added viethanol(5m1) under cooling. After stirring for 30 minutes, the solution was set aside overnight in a refrigerator.

The resulting crystals were filtered and washed with a mixture of water and acetone to give the title compound(2.5g, 67.6%).

Example 43.

2'-Benzyloxycarbonylphenyl trans-4-guanidinomethylcyclohexane- carboxyl ate hydrochloride monohydrate: 2'-BenzyloxyNcarbonylphenyl trans-4-guanidinomethylcyclo- hexanecarboxylate hydrochloride was recrystallized from acetone/ water(85/15), and dried at 400 C to give the title compound.

m.p. 96-1050 C.

NMR: Spyridine-d

1.0-3.4 12H)

5.25 2H)

5..4 2H)

S 7.3-9.1 14H) @6 0

Analysis: Found: C, 59.61, H, 6.57, N, 9.18, C1, 7.67 o C C. C23 H2 7 N304 HC1 H20 S requires: C, 59.54, H,.6.52, N, 9.06, C1, 7.64.

Water Determination(Karl Fischer):

Found: 3.98 C2 3H2 7N 304HC1 H0 requires: 3.88.

*e C Exaumle 44.

4'-Ethoxycarbonylrethylcarbonylphenyl trans-4-guanidinmthylcycloherane- SOO 6~ B

carboxylate hydrochloride:

A solution of trans-4-guanidinamethylcyclohexanecarboxylic acid

hydrochloride (9.4 etoxycarbonylethyl-p-hydroxybenzoate (9.0 g)

and dicyclohexylcaro diimide (9.1 g) in dimethylformmide (50 mi)

was stirred at room tenperature for 24 hours. Insoluble materials

were removed by filtration, and the filtrate was evaporated to dryness.

lb the residue was added water, and the solution was acidified with

hydrochloric acid and extracted with chloroform. The extract was

evaporated to dryness. The residue was recrystallized from methanol

to obtain the title ccmpound (10.6 g,

.,f er

1111.05 I- 1 1. j IIIII=

RESOLUTION TEST CHART MICROCOPY NATIONPJBUREAUOF 51,,qDARDS-1963-A

140-1450 C 1 xmax (anr 1735, 1745, 1760

NR: 6CD3 OD

0.9 3.1

4.25 2H)

4 9 2H) *SsS

7.'25, 8.1 4H) Se

to 0 4

.*st Ebampfe

4' -Diphenyl trans-4-guanidi thylcyclohexanecarboxylate hydrochloride:

LO A solition of trans-4-goaanidinorrethycyclohexanecarboxylic acid

methanesulfonate (1.0 g) and dimethyfornamide (0.5 in thionyl

4 chloride (5 mZ) was heated at 55-600 C for 2.5 hours. After being 00

cooled, the reaction mixture was washed with petroleum ether and

dissolved in chlorofonm. The resulting clear solution was added to

a solution of p-phenyl-phenol (0.69 g) in pyridine (5 my), and the

mixiure was stirred overnight, at roim temperature. The reaction

S mikture was evaporated, and to the residue was added water. The

solution was extracted with chloroform. The extract was concentrat-

ed.and chroatographed on a column filled with silica gel using

chlorofonVm/methanol as an eluent. The resulting crude product was

then tre .ted with ether/methanol/6N hydrochloric acid to give the

title copound.

Mcmple 46.

4-Diphenyl trans-4-guanidinrmethylcyclohexanecarboxylate hydrochloride:

A mixture of trans-4-guanidinormethylcyclohexanecarboxylic acid

hydrochloride (1.0 g) and thionyl chloride (5 m) was refluxed for

2 hours. After concentration, to the reaction mixture was added

chlproform (10 mn) The solution thus obtained was added to a 5e50

solution of p-phenylphenol (0.7 g) in pyridine (5 mi) and heated

at 40-50 0 C for 2 hours. Eb the ieaction mixture was added hydro-

chloric acid, and the resulting solution was agitated and purified

chramatographically to obtain the title compound.

Example 47. ese **so 4 -Carbcxymethylpheny1 trans-4-guanidinonethylcyclohexanecarboxylate 4 0O

0 hydrochloride:

A suspenioan of trans-4-guanidinonethylcyclohexanecarboxylic acid

hydrochloride (10.64 benzyl p-hydroxyphenylacetate (10.94 g) and

dicylohexylcarboxylicimide (11.18 g) in pyridine (40 mn) was

stirred at room tenperature for 40 hours. After removal of insoluble

materials by filtration, the filtrate was concentrated, and to the

residue was- added water. The solution was acidified with hydrochloric

fO acid to obtain precipitated materials, which precipitates were collected

by filtration and extracted with methanol. The extract obtained was

evaporated to dryness in vacuo. The oilyresidue was dissolved in

methanol acetic acid and hydrogenated over pd-c. After absorption of hydrogen, the catalyst was remrved by filtration, and the filtrate was concentrated. Ib the residue was added acetone to obtain crystals. The crystals were collected by filtration and recrystallized frnom ethanol to give the title corp)ound (5.4 g, 32.4%).

174-176.5 0C

I.R: (cmnl) 1720, 1750 (C4)

M AR:

0.9 3.1 12H)

3.6 2H) 3*

6.95 7.4 4H)

Example 48. *O 4 (2"-Benzyloxycarbnyl) phenyl trans-4-guanidinonethylcyclohexancarboxy- a*

Slate hydrochloride: 0U0O

Ethylchloroformate (0.65 g) was added to a suspension of carboxyethyl)phenyl trans-4-guanidinarmethylcyclohexanecarboxylate

hydrochloride (1.92 g) in pyridine (3 mi) and chloroform (10 mi) at -15 to

-10 0 C. After agitation of the mixture at the same tenperature for

1 hour, to the reaction mixture was added benzyl alcohol (1.08 g).

The resulting solution was stirred continuously for 1qminutes at

to--100 C. The reaction mixture was evaporated to dryness in vacuo,

and to the residue were water and hydrochloric acid. The solution

thus obtained was extracted with chloroform. The solvent was renoved

from the extract. To the residue was added water to give the title

compound.

Exrnple 49.

4 1-Carboxymethylphenyl trans-4-guanidinonethylcyclohexanecarboxylate

hydrochloride:

1) A solution of trans-4-guanidinornethylcyclohexanecarboxylic acid

hydrochloride (4.7 tert-butyl p-hydroxyphenyl acetate (5.5 g) and

dicyclohexylcarbodiinide (6.2 g) in dimethylformamide, (20 mk) was

stirred at roa temperature for 20 hours. The reaction mixture was

evaporated in.:--icuo, and to the residue was added chloroform. Insoluble

materials were removed by filtration, and the filtrate was washed

with water. The thus obtained chloroform layer was concentrated to

give '-tert-butoxycarbonyluethylphenyl trans-4-guanidinamethylcyclo-

hexanecarboxylate hydrochloride (6.9 g, 81.0%). see*

110 1250 C

2) A solution of 4' -terL--butoxycarbonyuethylphenyl trans--4-

guanidinriothylcyclohiexanecarboxylate hydrochloride (3.0 g) in acetic

acid (30 m9L) ard a 15% hydrogen chloride-acetic acid solution was

stirred.at room temperature for 4 hours. The reaction mixture was

evaporated to dryness in vacno to give the title caripound.

Examnple

2'-Benzyyoxycarbonylphenyl trans-4-~anidrmiethylcyclohexanecarboxyLate

hydrochloride:

1) trans-4-Guanidimuiothylcyclohexanecarboxylic acid chloride

hydrochloride:

A suspension of trans-4-guanidinamethy1cyclcbe-anecarboxylc acid

hydrochloride (1.2 g) and thionyl chloride (15 mk) was stirred at

ronom temperature for 3 hours. Thereafter, excess thionyl chloride

was renoved by distillation under reduced pressure. The residue was

washed with anhydrous ethyl ether to obtain trans-4-guanidinamethyl-

cyclohexanecarboxylic acid chloride hydrochloride (1.2 g) as colorless

Fowder.

IR nujol cmn1 *max 1790 (C=O)

2) 2' -Benzyloxycarbonnylphenyl trans-4-guanidinanethylcyclohexane-

carboxylate hydrochloride:

A solution of triethylamine (0.75 g) in anhydrous methylenechloride

(2 m9) was added to a suspension of trans-4-guanidinomethylcyclohexane-

carboxylic acid chloride hydrochloride (1.2 g) and benzyl salicylate

Volp 4 (1 in anhydrous methylenechloride (10 mi) at 0 0 C, and the resulting

mixture was stirred at 00 C for 10 hours. After distillation of the

solvent, the residue was washed with a saturated sodium hydrogen

carbonate solution to remove unreacted trans-4-guanidinorethylcyclo-

shexanecarboxylic acid, and to the resulting residue was added anhydrous

sodium sulfate. Then, the mixtunre was extracted with methylene chloride.

After distillation of the solvent, the residue was washed with ether

to renove unreacted benzyl salicylate. To the residue was added

isopropyl alcohol, and insoluble materials were then remnvad by

filtration. The solvent was reoved from the filtrate by distillation,

and the resulting residue was recrystallized from water to give

2 '-benzyloxycarboylphenyl trans-4-guanidinomnethylcyclohexanecarboxylate

hydrochloride monohydrate (320 mg) as colorless crystals.

Examrple 51.

2' -Benzyloxycarlbnnylphenyl trans-4-guanidir-&nthylcyclohexanecarboxylate

hy [rochioride monohydrate:

trans-4-uanidinonthylcyclohexanecarbxylic acid hydrochloride

(1.71 g) and trietbrftnine (0.74 g) were suspended in methylenechioride

mk), and to the supension was added isobutyloxycarhonylchloride

(1 g) at -51C. To the resulting inixture, 'hiAch had been stirred at -50c for 30 minutes, was added bc-nzyl salicylate Ilt (1.66 16 The resut- ).Tersl-

a /lng mixture was stirred for 2 hours under ice oooling and further b.~l

;stirred for 36 hours at roan temperature. After distillation of the solvent, the residue was washed with a satu-rated sodium hydrbgencarbonate

solution and extracted with methylenechloride. After distillation

of the solvent, the residue was washed with ether and purified by

thin-layer chromatography (n-butanol: acetic acid: water-4:1:1) to *00

give 2' -benzyoxycarbonyphenyl trans-4-guanidinc'n-ethylcyclohexane- carboxylate hydrochloride monohydrate (285 mg).

*ease: .0 0

Example 52.

Tablets; These contain the following materials in 220mg of a

film-coating tablet.

Compound 1

Cornstarch 100mg

Crystalline cellose

Magnesium stearate 1mg

Hydroxypropyl methyl cellose o

"LO Hydroxypropyl cellose 4mg

Total 220mg

Compound 1 is the same as defined above.

Granules: These contain the following materials in 1000mg of a granules.

Compound 1 100mg

Avicel 500mg

Cornstarch 400mg

Total 1000mg

Compound 1 is the same as defined above.

0 Having fully described the invention, it will be apparent to

one skilled in this art that many changes and modifications can be

made without departing from the spirit or scope of the invention.

Claims (1)

1. or:

   -69-

   THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:

   1. A compound of the formula:

   see* o HN -NHCH HN KD COOl H2 N NC 2 U-COR

   wherein R represents a N vanilyl, napthyl, pyridyl or a-tocopheryl 5O 0 A 0 R2 2 epesnt group, or a group of the formula, wherein R 2 represents

   a hydrogen atom, a lower alkoxy, formyl, lower alkanoyl or phenyl

   group, or a group of the fonrmula, -(Ca 2)n QO, wherein represents a hydrogen atm, a lower alkyl, phenyl, benzyl, anisyl 0*

   66:6or lower alkoxycarboanythethyl group, and n represents an integer .0 0 of 0 to 2, or a pharmnaceutically acceptable salt thereof.

   So** 002. A compound of the formula: *9000: 0 HN

   C-NHCH 2 COOR 1 H2N

   wherein R1 represents a vanilyl, naphthyl, pyridyl or a-tocophery R2 group, or a group of the fonmila wherein R2 represents

   a hydrogen atom, a lower alkoxy, formyl, Idwer alkanoyl or phenyl

   group, or a group of the formula -(CH2) nCDOR3, wherein R3 represents sip+ a hydrogen atom, or a lower alkyl, phenyl, benzyl, anisyl or lower alkoxycarxbonyliethyl group, and n represents an integer of 0 to 2, or a pharmaceutically acceptable salt thereof.

   3. A compound of the formula:

   S* @0 HN R

   00 -NHCH 22 2 -O

   H2N

   wherein R2 represents a hydrogen atom, a lower alkoxy, formyl, lower

   alkanoyl or phenyl group,or a group of the formula, -(CH )nCOR3 0 wherein R3 represents a hydrogen atom, or a lower alkyl, phenyl, benzyl,

   anisyl or lower alkoxycarbonylmethyl group,and n represents an 00 Iteger of 0 to 2, or a pharmaceutically acceptable salt thereof.

   4. A compound of the formula: 0( S*

   H JC-NHCH2-Q 0 0 ,(C2 n03

   HN

   wherein R3 represents a hydrogen atom, or a lower alkyl, phenyl, benzyl,

   anisyl or lower alkoxycarbonylmethyl group, and n represents an

   integer 0 to 2, or a pharmaceutically acceptable salt thereof.

   Vanilyl 4-guanidinomethylcyclohexanecarboxylate or a

   pharmaceutically acceptable salt thereof.

   -71-

   6. Naphthyl 4-guanidinomethylcyclohexanecarboxylate or a

   pharmaceutically acceptable salt thereof.

   7. Pyridyl 4-guanidinomethylcyclohexanecarboxylate or a

   pharmaceutically acceptable salt thereof.

   8. a-Tocopheryl 4-guanidinomethylcyclohexanecarboxylate ester

   or a pharmaceutically acceptable salt thereof.

   -Benyloxycarbonyl )phenyl trans-4-guanidinomethylcyclonexane-

   ri_ carboxylate hydrochloride.

   1. (2'-Benzyloxycarbonvl)phenyl trans-4-guanidinomethylcycl o -xane-

   carbcxylate hydrochloride monohydrate.

   11. A process for producing a compound of the formula:

   -NHCH2-- COORn

   COR2 r wherein R1 represents a vanilyl, napthyl, pyridyl, a-tocopheryl

   group, or a group of the formula, wherein R represents a hydrogen atom, a lower alkoxy, formyl, lowr

   alkanoyl or phenyl groap, or a group of the formla, -(CU 2 naOQP- whe~rein R3 represents a hydrogen atom or a lower alkyl, phenyl, benzyl, anisyl or lower alkoxycarbonylmethyl group, and n represents an integer of 0 to 2, which comprises reacting 4-guanidinomethylcyclohexanecarboxylic acid or a reactive derivative thereof with a compound of the formula,

   *006 o- OH

   *.wherein R1 represents a vanilyl, naphthyl, pyridyl or a-tooopheny1

   group, or a group of the formla, \,wherein R'2 represents a hydrogen atomn, a lower alkoxy, fontmrl, lower alkanoyl or pbenyl group, or a group of the formnula (CH 2 nCC)R, wherein e~g.

   R represents a lower alkyl, phenyl, benzyl, anisyl(lower alkoxy- carbonylmethyl -gr--buty14 group, and n aersentsa integer of 0 to 2.

   12. The process of Claim 11, wherein said benzyl, anisyl, lower

   alkoxycarbonyLmethYl or t-butyl group is replaced by a

   13. The process of Claim 11 or 12,wherein said compound is further treated with a pharmaceutically acceptable acid to form the corresponding salt thereof.

   14. A process for producing a compound of the formula,

   HNi' COOR' ~NHCH 2 .Q.a Coo (CH 2 )n 3

   H2 wherein R' represents a lower alkyl, phenyl, benzyl, anisyl or lower alkoxycarbonylmethyl group, and n represents an integer of 0 to 2, which comprises reacting a compound of the formula,

   S C-NHCH 2 -nCH 2 COOH

   HN 2

   wherein n is the same as defined above, or a reactive derivative o 0.

   thereof with a compound of the formula,

   R' OH 33 S(

   wherein R is the same as defined above.

   15. An anti-ulcer agent which comprises a compound of the formula, H *H NHCH2 -3- (CH 2)n COOR 3

   H2 N.

   wherein R3 represents a hydrogen atom or a lower alkyl, phenyl, benzyl,

   anisyl or lower alkoxycarbonylmethyl group, and n represents an integer

   of 0 to 2,or a pharmaceutically acceptable salt thereof.

   16. The anti-ulcer agent of Claim 15 which is (2'-benzyloxycarbonyl)-

   phenyl trans-4-guanidinomethylcyclohexanecarboxylate hydrochloride.

   74.-

   17. The anti-ulcer agent of Claim 15 which is (2'-benzyloxycarbonyl)phenyl trans-4-guanidinomethylcyclohexanecarboxyilate hydrochloride monohydrate.

   DATED this 10th day of September, 1980

   egg. 0 NIPPON CHEMIPHAR CO., LTD. and gte. TEIKOKU CHEMICAL INDUSTRY CO., LTD. me S *O 0* Patent attorneys for the applicant OS OS, 0 '0 0

   00 0 F.B. RICE CC S S *5

   go go ego.

   egg.. C *060

   Ce 0 *g

   egee gg C* C

   6L